Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine originally named for its ability to transform normal fibroblasts to cells capable of anchorage-independent growth. The TGF-betas, produced primarily by hematopoietic and tumor cells, can regulate, i.e., stimulate or inhibit, the growth and differentiation of cells from a variety of both normal and neoplastic tissue origins (Sporn et al., Science, 233: 532 (1986)) and stimulate the formation and elaboration of various stromal elements. For a general review of TGF-beta and its actions, see Sporn et al., J. Cell Biol., 105: 1039-1045 (1987) and Sporn and Roberts, Nature, 332: 217-219 (1988).
They are known to be involved in many proliferative and non-proliferative cellular processes such as cell proliferation and differentiation, embryonic development, extracellular matrix formation, bone development, wound healing, hematopoiesis, and immune and inflammatory responses. Pircher et al, Biochem. Biophys. Res. Commun., 136: 30-37 (1986); Wakefield et al., Growth Factors, 1: 203-218 (1989); Roberts and Sporn, pp 419-472 in Handbook of Experimental Pharmacology eds M. B. Sporn & A. B. Roberts (Springer, Heidelberg, 1990); Massague et al., Annual Rev. Cell Biol., 6: 597-646 (1990); Singer and Clark, New Eng. J. Med., 341: 738-745 (1999). Also, TGF-beta is used in the treatment and prevention of diseases of the intestinal mucosa. WO 2001/24813.
Of particular interest from an immunological viewpoint are the potent immunosuppressive activities of TGF-beta, which include lymphokine-activated killer (LAK) and cytotoxic T lymphocyte (CTL) inhibition (Ranges et al., J. Exp. Med., 166: 991 (1987), Espevik et al., J. Immunol., 140: 2312 (1988), Grimm et al., Cancer Immunol. Immunother., 27: 53 (1988), Kasid et al., J. Immunol., 141: 690 (1988), Mule et al., Cancer Immunol. Immunother., 26: 95 (1988)), depressed B cell lymphopoiesis and kappa light-chain expression (Lee et al., J. Exp. Med., 166: 1290 (1987)), negative regulation of hematopoiesis (Hino et al., Br. J. Haematol., 70: 143 (1988), Sing et al., Blood, 72: 1504 (1988)), down-regulation of HLA-DR expression on tumor cells (Czarniecki et al., J. Immunol., 140: 4217 (1988), Zuber et al., Eur. J. Immunol., 18: 1623 (1988)), and inhibition of the proliferation of antigen-activated B lymphocytes in response to B-cell growth factor (Petit-Koskas et al., Eur. J. Immunol., 18: 111 (1988)). The observation that many human tumors (deMartin et al., EMBO J., 6: 3673 (1987), Kuppner et al., Int. J. Cancer, 42: 562 (1988)) and many tumor cell lines (Derynck et al., Cancer Res., 47: 707 (1987), Roberts et al., Br. J. Cancer, 57: 594 (1988)) produce TGF-beta suggests a possible mechanism for those tumors to evade normal immunological surveillance. This negative immunomodulation, coupled with the observations that certain transformed cell lines have lost the ability to respond to TGF-beta in an autocrine fashion (Wakefield et al., J. Cell Biol., 105: 965 (1987), McMahon et al., Cancer Res., 46: 4665 (1986)), and that TGF-beta stimulates stroma formation, and decreases immune surveillance of the tumor, suggests attractive models for neoplasm deregulation and proliferation (Roberts et al., Br. J. Cancer, supra). 
In addition, U.S. Pat. Nos. 5,824,297 and 5,262,319 disclose a method for inhibiting cytotoxic poisoning of normal cells by administering thereto a TGF-beta such as TGF-beta3.
There are at least five forms of TGF-beta currently identified, TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, and TGF-beta5. Suitable methods are known for purifying this family of TGF-betas from various species such as human, mouse, green monkey, pig, bovine, chick, and frog, and from various body sources such as bone, platelets, or placenta, for producing it in recombinant cell culture, and for determining its activity. See, for example, Derynck et al., Nature, 316: 701-705 (1985); European Pat. Pub. Nos. 200,341 published Dec. 10, 1986, 169,016 published Jan. 22, 1986, 268,561 published May 25, 1988, and 267,463 published May 18, 1988; U.S. Pat. No. 4,774,322; Cheifetz et al, Cell, 48: 409-415 (1987); Jakowlew et al., Molecular Endocrin., 2: 747-755 (1988); Dijke et al., Proc. Natl. Acad. Sci. (U.S.A.), 85: 4715-4719 (1988); Derynck et al., J. Biol. Chem. 261: 43774379 (1986); Sharples et al., DNA, 6: 239-244 (1987); Derynck et al., Nucl. Acids. Res., 15: 3188-3189(1987); Deryncketal., Nucl. Acids. Res., 15: 3187(1987); Derynck et al., EMBO J., 7: 3737-3743 (1988)); Seyedin et al., J. Biol. Chem. 261: 5693-5695 (1986); Madisen et al., DNA 7: 1-8 (1988); and Hanks et al., Proc. Natl. Acad. Sci. (U.S.A.), 85: 79-82 (1988), the entire contents of these publications being expressly incorporated by reference.
The activated form of TGF-beta1 is a homodimer formed by dimerization of the carboxy-terminal 112 amino acids of a 390-amino-acid precursor (Derynck et al., Nature, supra). TGF-beta2 has a precursor form of 414 amino acids and is also processed to a homodimer from the carboxy-terminal 112 amino acids that shares approximately 70% homology with the active form of TGF-beta1 (Marquardt et al., J. Biol. Chem. 262: 12127 (1987)). TGF-beta2 has been purified from porcine platelets (Seyedin et al., J. Biol. Chem., 262: 1946-1949 (1987)) and human glioblastoma cells (Wrann et al., EMBO J., 6: 1633 (1987)), and recombinant human TGF-beta2 has been cloned (deMartin et at, supra). Recombinant TGF-beta1 has been cloned (Derynck et al., Nature supra) and expressed in Chinese hamster ovary cells (Gentry et al., Mol. Cell. Biol., 7: 3418-3427 (1987)). See U.S. Pat. Nos. 4,774,322; 4,843,063; and 4,848,063 regarding CIF-A and CIF-B, now recognized as TGF-beta1 and 2, respectively. Ellingsworth et al., J. Biol. Chem., 261: 12362-12367 (1986). Even though there are 14 amino acid differences in the first 36 amino acid residues of the two forms (TGF-beta1 and TGF-beta2), their biological activities are similar. Cheifetz et al., Cell, 48: 409-415 (1987); Seyedin et al., J. Biol. Chem., 262: supra.
TGF-beta3, TGF-beta4, and TGF-beta5, which are the most recently discovered forms of TGF-beta, were identified by screening cDNA libraries. None of these three putative proteins has been isolated from natural sources, although Northern blots demonstrate expression of the corresponding mRNAs. Human and porcine TGF-beta3 have been cloned and are described as homodimers and expressed in Chinese hamster ovary cells (Derynck et al., EMBO J., 7: 3737-3743 (1988), ten Dijke et al., Proc. Natl. Acad. Sci. USA, 85: 4715 (1988); U.S. Pat. No. 4,886,747). See also WO 1992/00318 regarding TGF-beta3 proteins and antibodies thereto. TGF-beta1 differs from TGF-beta2 by 27 mainly conservative changes and from TGF-beta3 by 22 mainly conservative changes. These differences have been related to the 3-D structure. Schlunegger and Grutter, Nature, 358: 430-434 (1992).
TGF-beta4 and TGF-beta5 were cloned from a chicken chondrocyte cDNA library (Jakowlew et al., Molec. Endocrinol., 2: 1186-1195 (1988)) and from a frog oocyte cDNA library, respectively. The frog oocyte cDNA library can be screened using a probe derived from one or more sequences of another type of TGF-beta. TGF-beta4 mRNA is detectable in chick embryo chondrocytes, but is far less abundant than TGF-beta3 mRNA in developing embryos or in chick embryo fibroblasts. TGF-beta5 mRNA is expressed in frog embryos beyond the neurula state and in Xenopus tadpole (XTC) cells.
The recombinant production of TGF-beta1, TGF-beta2, and TGF-beta3 is described in U.S. Pat. Nos. 5,061,786; 5,268,455 and 5,801,231. See also U.S. Pat. No.5,120,535 on a TGF-beta2 used for treating hormonally responsive carcinoma and for production of antibodies. The heterodimer of TGF-beta1 and TGF-beta2, called TGF-beta1.2, has been identified and its uses demonstrated, as disclosed in U.S. Pat. Nos. 4,931,548 and 5,304,541, the latter also disclosing an antibody thereto. WO 1990/00900, filed 20 Jul. 1989, discloses treatment of inflammatory disorders with homodimeric TGF-beta1 and -beta2, and the heterodimer TGF-beta1.2. U.S. Pat. No. 5,462,925 discloses a heterodimer of TGF-beta2 and TGF-beta3. U.S. Pat. No. 5,780,436 discloses small peptide mimics of TGF-beta.
Increased levels of TGF-beta activity are involved in a large number of pathologic conditions, including, but not limited to, the following: (i) fibrosis, scarring, and adhesion during wound healing; (ii) fibrotic diseases of the lungs, liver, and kidneys; (iii) atherosclerosis and arteriosclerosis; (iv) certain types of cancer including cancer of the prostate, neuroendocrine tumors of the digestive system, cancer of the cervix, glioblastomas, and gastric cancer; (v) angiopathy, vasculopathy, nephropathy; (vi) systemic sclerosis; (vii) viral infection such as hepatitis C and HIV; and (viii) immunological and inflammatory disorders and deficiencies such as rheumatoid arthritis. The modulation of immune and inflammatory responses by TGF-betas includes: (i) inhibition of proliferation of all T-cell subsets; (ii) inhibitory effects on proliferation and function of B lymphocytes; (iii) down-regulation of natural-killer cell activity and the T-cell response; (iv) regulation of cytokine production by immune cells; (v) regulation of macrophage function; and (vi) leukocyte recruitment and activation.
As to cancer specifically, members of the TGF-beta family are known to have a number of biological activities related to tumorigenesis (including angiogenesis) and metastasis. TGF-beta inhibits the proliferation of many cell types including capillary endothelial cells and smooth muscle cells. TGF-beta downregulates integrin expression (alpha1beta1, alpha2beta1, and alphavbeta3 involved in endothelial cell migration). Integrins are involved in the migration of all cells, including metastatic ones. TGF-beta downregulates matrix metalloproteinase expression needed for both angiogenesis and metastasis. TGF-beta induces plasminogen activator inhibitor, which inhibits a proteinase cascade needed for angiogenesis and metastasis. TGF-beta induces normal cells to inhibit transformed cells. See, e.g., Yingling et al., Nature Reviews, 3 (12): 1011-1022 (2004), which discloses that deregulation of TGF-beta has been implicated in the pathogenesis of a variety of diseases, including cancer and fibrosis, and presents the rationale for evaluating TGF-beta signaling inhibitors as cancer therapeutics, biomarkers/diagnostics, the structures of small-molecule inhibitors that are in development, and the targeted drug discovery model that is being applied to their development. Early detection of cancer is very important (Ruth et al., Nature Reviews Cancer, 3: 243-252 (2003)), and the pathogenesis of cancer metastasis is being studied. Fidler, Nature Reviews Cancer, 3: 453-458 (2003).
TGF-beta has emerged to be a major modulator of angiogenesis by regulating endothelial cell proliferation, migration, extracellular matrix (ECM) metabolism, and the expression of adhesion molecules. It is a potent growth inhibitor of normal mammary epithelial cells and a number of breast cancer cell lines. TGF-beta appears to exert pleiotropic effects in the oncogenesis of breast cancers in a contextual manner, i.e., it suppresses tumorigenesis at an early stage by direct inhibition of angiogenesis and tumor cell growth. However, over-production of TGF beta by an advanced tumor may accelerate disease progression through indirect stimulation of angiogenesis and immune suppression. The cell membrane antigen CD105 (endoglin) binds TGF beta1 and TGF beta3 and is preferentially expressed in angiogenic vascular endothelial cells. The reduction of CD105 levels in HUVEC leads to in vitro angiogenesis inhibition and massive cell mortality in the presence of TGF-beta1. CD105 null mice die in utero with impaired vasculature, indicating the pivotal role of CD105 in vascular development. Li et al., Microsc. Res. Tech., 52:437-449 (2001). Abnormal angiogenesis but intact hematopoietic potential has been observed in TGF-beta type I receptor-deficient mice. Larsson et al., EMBO J., 20 (7): 1663-1673 (2001). Further, TGF-beta receptor type II deficiency resulted in defects of yolk sac hematopoiesis and vasculogenesis. Oshima et al., Developmental Biology, 179 (1): 297-302 (1996). Also, heart and liver defects and reduced transforming growth factor beta 2 sensitivity were observed in TGF-beta type III receptor-deficient embryos. Stenvers et al., Mol. Cell. Biol., 23 (12): 43714385 (2003). Further, targeted disruption of the mouse TGF-beta1 gene resulted in multifocal inflammatory disease. Shull et al., Nature, 359 (6397): 693-699 (1992). Early-onset multifocal inflammation in the TGF-beta1-null mouse was found to be lymphocyte mediated. Diebold et al., Proc. Natl. Acad. Sci. (USA), 92 (26): 12215-12219 (1995).
The most important non-proliferative function of TGF-betas is in enhancing the formation of extracellular matrices. Although this is achieved primarily through the increased transcription of both collagen and fibronectin, the inhibition of the proteases from degrading the matrix also contributes to its stability. Degradation of the extracellular matrix is inhibited by the decrease in the secretion of the proteases themselves and the simultaneous increase in the levels of protease inhibitors.
WO 1984/001106 describes TGF-beta1 and its use for the promotion of cell proliferation and tissue repair, wound healing, and treatment of traumata. U.S. Pat. No. 4,806,523 discloses that TGF-beta1and TGF-beta2 both possess anti-inflammatory activity and are inhibitors of mitogen-stimulated T-cell proliferation and B-cell activation. It also reports that TGF-beta is localized in centers of hematopoiesis and lymphopoiesis and that TGF-beta may, therefore, be useful for treating indications associated with malfunction or dysfunction of hematopoiesis or lymphopoiesis.
TGF-beta2 has been shown to be the predominant isoform of TGF-beta in the neural retina, retinal pigment epithelium-choroid and vitreous of the human eye (Pfeffer et al., Exp. Eye Res., 59: 323-333 (1994)) and found in human aqueous humour in specimens from eyes undergoing cataract extraction with intraocular lens implantation. Jampel et al., Current Eye Research, 9: 963-969 (1990). Non-transformed human retinal pigment epithelial cells predominantly secrete TGF-beta2. Kvanta, Ophthalmic Res., 26: 361-367 (1994).
Other diseases that have potential for treatment with antibodies against TGF-beta include adult respiratory distress syndrome, cirrhosis of the liver, post-myocardial infarction, post-angioplasty restenosis, keloid scars, and scleroderma. The increased level of expression of TGF-beta2 in osteoporosis (Erlenbacher et al. J. Cell Biol., 132: 195-210 (1996)) means that this is a disease potentially treatable by antibodies directed against TGF-beta2.
Because of the involvement of TGF-beta in a large number of serious pathological conditions, there is considerable interest in developing inhibitors of TGF-beta. Many of the proposals for TGF-beta inhibitors have involved antibodies.
It is a demanding task to isolate an antibody fragment specific for TGF-beta of the same species. Animals do not normally produce antibodies to self-antigens, a phenomenon called tolerance (Nossal, Science, 245: 147-153 (1989). In general, vaccination with a self-antigen does not result in production of circulating antibodies. It is therefore difficult to raise human antibodies to human self-antigens. There are also, in addition, ethical problems in vaccinating humans. In relation to the raising of non-human antibodies specific for TGF-beta, there are a number of problems. TGF-beta is an immunosuppressive molecule and further, there is strong conservation of sequence between human and mouse TGF-beta molecules. Mouse and human TGF-beta1 only differ by one amino acid residue, an alanine (human)-to-serine (mouse) change at a buried residue. Derynck et al., J. Biol. Chem., 261: 4377-4379 (1986). Mouse and human TGF-beta2 only differ at three residues; residue 59 (T mouse, S human); residue 60 (K mouse, R human), and residue 94 (N mouse; K human). This makes it difficult to raise antibodies in mice against human TGF-beta. Further, any antibodies raised may only be directed against a restricted set of epitopes.
Monoclonal antibodies against TGF-beta have been produced by immunizing chickens and immortalizing B cells, used for, e.g. diagnosis and passive treatment of disease as described in U.S. Pat. No. 6,143,559.
Polyclonal antibodies binding to human TGF-beta1 and human TGF-beta2 against both neutralizing and non-neutralizing epitopes have been raised in rabbits (Danielpour et al., Growth Factors, 2: 61-71 (1989); Roberts et al. Growth Factors, 3: 277-286 (1990)), chickens (R&D Systerrs, Minneapolis) and turkeys (Danielpour et al., J. Cell Physiol., 138: 79-86 (1989); Danielpour and Sporn, J. Cell Biochem., 13B: 84 (1989)).
Peptides representing partial or complete TGF-beta sequences have also been used as immunogens to raise neutralizing polyclonal antisera in rabbits. Ellingsworth et al., J. Biol. Chem. 261: 12362 (1986); Ellingsworth et al., Cell. Immunol., 114: 41 (1988); Border et al. Nature 346: 371-374 (1990); Flanders, Biochemistry 27: 739-746 (1988); Flanders et al., Growth Factors 3: 45-52 (1990); Flanders et al., Development, 113: 183-191 (1991). In addition, there have been limited reports of isolation of mouse monoclonal antibodies against TGF-beta. Following immunization with bovine TGF-beta2 (identical to human TGF-beta2), three non-neutralizing monoclonal antibodies were isolated that are specific for TGF-beta2 and one neutralizing antibody that is specific for TGF-beta1 and TGF-beta2. Dasch et al., J. Immunol., 142: 1536-1541 (1989). In another report, following immunization with human TGF-beta1, neutralizing antibodies were isolated that were either specific for TGF-beta1 or cross-reacted with TGF-beta 1, TGF-beta2 and TGF-beta3. Lucas et al., J. Immunol., 145: 1415-1422 (1990). Polyclonal antisera to human and porcine TGF-beta (Keski-Oja et al., Cancer Res., 47: 6451-6458 (1987)) and to porcine TGF-beta2 (Rosa et al., Science, 239: 783-785 (1988)) have been shown to neutralize the biological activity of TGF-beta1 and TGF-beta2, respectively. Rabbit anti-TGF-beta serum is described in Roberts et al., Proc. Natl. Acad. Sci. USA, 83: 4167-4171 (1986). In addition, RIAs against TGF-beta1 using rabbit antiserum have been established to quantitate the released protein during platelet aggregation. Assoian and Sporn, J. Cell Biol., 102: 12178-1223 (1986).
A neutralizing mouse monoclonal antibody that binds both TGF-beta2 and TGF-beta3 isoforms is available commercially from Genzyme Diagnostics. A mouse monoclonal antibody directed against human TGF-beta1 is available from R&D Systems. This antibody only weakly neutralizes TGF-beta1 in a neutralization assay. Neutralizing mouse monoclonal antibodies have also been generated from mice immunized with human TGF-beta1 peptides comprising amino acid positions 48 to 60 (antibody reactive with TGF-beta1, TGF-beta2 and TGF-beta3) and amino acid positions 86 to 101 (antibody specific for TGF-beta1). Hoefer and Anderer, Cancer Immunol. Immunother., 41: 302-308 (1995).
Phage antibody technology (WO 1992/01047; WO 1993/19172; WO 1992/20791; WO 1993/06213; and WO 1993/11236) offers the ability to isolate directly human antibodies against human TGF-beta. The isolation of anti-self antibodies from antibody segment repertoires displayed on phage has been described. Griffiths et al., EMBO J., 12: 725-734 (1993); Nissim et al. EMBO J., 13: 692-698 (1994); Griffiths et al. 13: 3245-3260 (1994); Barbas et al., Proc. Natl. Acad. Sci. USA, 90: 10003-13007 (1993); and WO 1993/11236. In addition, Tempest et al., Immunotechnology, 2: 306 (1996) describes human antibodies specific for human TGF-beta derived from phage display libraries.
WO 1997/13844 discloses the isolation of human antibodies specific for human TGF-beta1 and human antibodies specific for human TGF-beta2. It describes antibodies with the 31 G9 VH domain and variants of the domain, more specifically, the antibody CS37 that comprises the 31G9 VH domain together with the CS37 VL and variants of this domain, including antibodies that: (i) compete in ELISA with CS37 for binding to TGF-beta 1, (ii) bind TGF-beta1 preferentially with respect to TGF-beta 3, and (iii) neutralize TGF-beta1.
U.S. Pat. No. 6,492,497 is based on identification of antibodies that are related to CS37, but that have unexpectedly advantageous properties with respect to binding and neutralization of TGF-beta1. They do not bind to, or neutralize, TGF-beta2 or TGF-beta3. The epitope for these antibodies lies in the C-terminal region of TGF-beta1 (residues 83-112) and includes the loop consisting of residues 92-98 of TGF-beta1, also known as finger 2, a region that has been identified as interacting with the receptor for TGF-beta.
A monoclonal antibody against human TGF-beta-1 that is highly specific and can be used for tumor diagnosis and for affinity chromatography is disclosed by JP 95068278 B2 published Jul. 26, 1995.
Use of TGF-beta and its antagonists for modulating blood pressure, and for treating hypertension and hypotension, respectively, is disclosed in WO 1991/19513.
WO 1991/15223 discloses a purified respiratory burst suppression factor that may be incubated with turkey anti-TGF-beta antibody that specifically binds TGF-beta1. The antibody completely neutralized the activity of TGF-beta1 on activated macrophages, but had no effect on the activity of the respiratory burst suppression factor on the macrophages.
Suppressing TGF-beta activity and extracellular matrix accumulation in diagnosis and treatment of fibrotic diseases such as glomerulonephritis by contact with an ECM-producing activity suppressor, such as anti-TGF-beta antibody, is disclosed in WO 1991/04748 and WO 1993/10808. Antibodies against a linear peptide from TGF-beta, and cells producing the antibodies are also disclosed.
U.S. Pat. No. 5,888,705 discloses a method of inducing the proliferation of human adult pancreatic cells or the differentiation thereof by contacting primary cultures of such cells with hepatocyte growth factor alone or in combination with anti-TGF-beta antibodies.
WO 2001/66140 discloses the use of TGF-beta antagonists such as antibodies to treat or prevent loss of renal function.
WO 2000/40227 discloses methods for treating conditions associated with the accumulation of excess extracellular matrix using agents that inhibit TGF-beta such as antibodies.
Antibodies to TGF-beta are disclosed as ameliorating tubular apoptosis in unilateral ureteral obstruction, in Miyajima et al., Kidney International, 58: 2301-2313 (2000).
Long-term prevention of renal insufficiency, excess matrix gene expression, and glomerular mesangial matrix expansion by treatment with monoclonal anti-TGF-beta antibody in db/db diabetic mice is disclosed in Ziyadeh et al., Proc. Natl. Acad. Sci. USA, 97 (14): 8015-8020 (2000).
Favorable treatment outcome with neutralizing anti-TGF-beta antibodies in experimental diabetic kidney disease is disclosed in Han and Ziyadeh, Peritoneal dialysis international, 19 Suppl 2: S234-237 (1999). TGF-beta was found to be a key mediator in hyperglycemia and diabetic kidney disease. Sharma and Ziyadeh, Diabetes, 44 (10) p1139-46 (1995). Use of TGF-beta in diabetic nephropathy is disclosed in Border et al., Diabetes Metab. Rev., 12/4: 309-339 (1996).
U.S. Pat. No. 5,662,904 describes a composition for use in treating wounds to inhibit scar tissue formation. An exemplary such composition has growth-factor-neutralizing antibody, such as antibodies to TGF-beta1, TGF-beta2, and PDGF.
U.S. Pat. No. 5,972,335 discloses compositions comprising at least two antibodies for use in promoting wound healing of fibrotic disorders, where the first antibody is specific for a single epitope on TGF beta1 and the second antibody is specific for a single epitope on TGF beta2.
U.S. Pat. No. 5,958,411 discloses methods for treating a CNS pathology by administering neutralizing anti-TGF-beta antibodies.
U.S. Pat. No. 5,616,561 describes a method for treating tissue damage caused by radiation using a TGF-beta antagonist such as antibodies.
U.S. Pat. No. 6,500,920 discloses a peptide of 10-25 amino acids comprising amino acids 49-58 of a TGF-beta2, wherein the peptide is capable of inhibiting specific binding of a TGF-beta to a TGF-beta receptor on a cell.
U.S. Pat. Appln. No. 2002/0176858 and U.S. Pat. Nos. 5,693,607; 6,419,928; 6,090,383; 5,783,185; 5,772,998; and 5,571,714, as well as EP 489,062; 557,418; and 669,833, as well as WO 1992/08480; 1994/09815; and 1994/18991 disclose monoclonal antibodies to TGF-beta, including ones that neutralize the activity of TGF-beta1 and TGF-beta2, and their use in therapeutic applications for treating indications where there is an overproduction of TGF-beta (e.g., acute liver injury, interstitial lung fibrosis, liver cirrhosis, chronic hepatic fibrosis, and fibrotic skin disorders such as scleroderma) and for diagnosing or treating malignancies (e.g., sarcomas and melanomas) and metastatic cancers.
New antibodies for treating disorders associated with TGF-beta3, e.g., osteoporosis, AIDS, cancer, etc., are disclosed in WO 1992/00330 and U.S. Pat. No. 5,262,319. Such antibodies bind to human TGF-beta3 and exhibit no cross-reactivity with TGF-beta1 and beta2.
U.S. Pat. No. 6,509,318 discloses a family of small peptides found to be inhibitory to TGF-beta activity for uses such as scar tissue inhibition during wound healing.
Use of a compound (e.g. an antibody) that can inhibit the biological activity of TGF-beta on pre-damaged neurons for treating cerebral disorders, e.g. cerebral ischemia, is disclosed in WO 2000/13705.
A monoclonal antibody recognizing all three isoforms of TGF-beta that can inhibit the biological activity of TGF-beta on pre-damaged neurons, useful for treating cerebral disorders, is disclosed in WO 2000/54804. Such antibody was used to neutralize endogenous TGF-beta during the main period of ontogenetic cell death of ciliary ganglia (CG) and dorsal root ganglia (DRG) as well as spinal motoneurons in chick embryos.
Diagnosing and predicting the likelihood of development of tamoxifen-sensitive or tamoxifen-resistant breast cancer using an antibody specific to angiogenic factors or receptors, such as an antibody specific to TGF-beta3, is disclosed in WO 2000/34788.
EP 945464 B1 discloses specific binding members for human TGF-beta, that is, specific binding members comprising human antibody-antigen binding domains specific for human TGF-beta that bind specifically isoforms TGF-beta2 and TGF-beta1, or both, preferentially compared with TGF-beta3. Specific binding members may be isolated and utilized in the treatment of disease, particularly fibrotic disease and also immune/inflammatory diseases.
Antibodies against TGF-beta have been shown to be effective in the treatment of glomerulonephritis (Border et al., Diabetes Metab. Rev., supra); neural scarring (Logan et al., Eur. J. Neurosci., 6: 355-363 (1994); WO 1993/19783); dermal scarring (Shah et al., Lancet, 339: 213-214 (1992); Shah et al., J. Cell Science, 107: 1137-1157 (1994); Shah et al., J. Cell Science, 985-1002 (1995); WO1992/17206); lung fibrosis (Giri et al., Thorax, 48: 959-966 (1993)); arterial injury (Wolf et al., J. Clin. Invest., 93: 1172-1178 (1994)); and rheumatoid arthritis (Wahl et al., J. Exp. Medicine, 177: 225-230 (1993)). It has been suggested that TGF-beta3 acts antagonistically to TGF-beta1 and TGF-beta2 in dermal scarring (Shah et al., 1995 supra).
Arteaga et al., J. Clin. Invest., 92: 2569-2576 (1993) discloses that anti-TGF-beta antibodies inhibit breast cancer cell tumorigenicity and increase mouse spleen natural killer cell activity.
Anti-fibrotic agents for wound healing and treatment of fibrotic disorders, including anti-TGF-betas, are described in WO 1993/19769.
Specific sequences of anti-TGF-beta2 are described in EP 853,661B1.
Other applications where antibodies against TGF-beta have shown promise of therapeutic efficacy include the use of antibodies against TGF-beta for the treatment of eye diseases involving ocular fibrosis, including proliferative retinopathy (Pena et al., Invest. Ophthalmology. Vis. Sci., 35: 2804-2808 (1994)), prevention of cataracts (WO 1995/13827), retinal detachment, and post glaucoma drainage surgery (Khaw et al., Eye, 8: 188-195 (1994)). Connor et al., J. Clin., Invest. 83: 1661-1666 (1989) showed that much higher levels of TGF-beta2 were present in vitreous aspirates from patients with intraocular fibrosis associated with proliferative retinopathy compared with patients with uncomplicated retinal detachment without ocular fibrosis and that the biological activity of this TGF-beta2 could be neutralized with antibodies directed against TGF-beta2.
The use of antibodies against TGF-beta for the treatment of diseases has been the subject of patent applications for fibrotic disease (WO 1991/04748); macrophage-deficiency diseases (WO 1993/14782); macrophage pathogen infections (WO 1993/17708; U.S. Pat. No. 5,730,976); and vascular disorders (WO 1993/21945).
A TGF-beta antibody-treated stem cell composition capable of survival for 14 days in vitro or ex vivo, and rapid in vivo hematopoietic system repopulation are described in WO 2000/43499.
Scrip 2580 p14, Oct. 04, 2000 reported that Cambridge Antibody Technology (CAT) and Genzyme were working together to develop human monoclonal antibodies against TGF-beta. CAT has two fully human TGF-beta antibodies, CAT-152 and CAT-192, and Genzyme has 1D11, a murine pan-specific monoclonal antibody that neutralizes TGF-beta1, TGF-beta2 and TGF-beta3 and is being evaluated as a potential therapeutic for diffuse scleroderma. CAT was to develop a human analogue of 1D11 using its phage display technology. Several other clinical indications for anti-TGF-beta treatment, including ophthalmic indications, post-surgical scarring, fibrosis of major organs, such as the lungs, kidneys and liver, and certain cancers, will also be considered as well as treatment of malignant brain tumors by inhibiting the growth of TGF-beta2. CAT-152 (anti-TGF-beta2) is in Phase II trials to prevent post-operative scarring in patients undergoing surgery for glaucoma, and CAT-192 (anti-TGF-beta1) has completed Phase I trials. See also “Trends in Antibody Research: The Monoclonal Elite” by Tim Searle, Bioventure-View 1510 pl4, Oct. 1,2000.
A method for quantifying TGF-beta using anti-TGF-beta antibody is disclosed in WO 1995/19987. A new assay for determining active TGF-beta in a sample using eukaryotic cells that contain a TGF-beta-responsive expression vector is described in WO 2000/00641. This assay includes one for determining the levels of TGF-beta isoforms in a sample, wherein cryosections are pre-incubated with anti-TGF-beta isoform neutralizing antibodies. TGF-beta immunoassays using TGF-beta antibodies are described, for example, in JP 2126157 and JP 92041307 B published Jul. 7, 1992.
Darland and D'Amore, J. Clin. Invest., 103: 157-158 (1999) discloses that vessel development proceeds from a stage of growth-factor dependence where loss of a survival factor leads to apoptosis. Vessel stabilization is marked by investment with mural cells, local activation of TGF-beta, and basement membrane production. It poses several questions regarding what is the role of growth factors in the adult vascular, including VEGF and TGF-beta. Benjamin et al., J. Clin. Invest., 103: 159-165 (1999) discloses selective ablation of immature blood vessels in established human tumors follows VEGF withdrawal.
Methods of making chimeric and humanized antibodies are described in, and other references in this area include, for example, U.S. Pat. No. 6,235,883 on fully human monoclonal antibodies against human epidermal growth factor receptor; EP 184187 on a mouse-human chimeric antibody; EP 844,306 on a method of making antibodies recombinantly using phage technology; U.S. Pat. No. 5,859,205 on preparing CDR-grafted antibodies, preferably humanized antibodies, having non-human donor and human acceptor frameworks, EP 120,694; EP 125,023; EP 171,496; EP 173,494; EP 239,400; WO 1989/07452; WO 1990/07861; and WO 1986/01533 on humanization techniques; U.S. Application No. 2003/0039649 on superhumanized antibodies; U.S. Application No. 2003/0039645 on humanized antibodies with specificity for human TNF-alpha; EP 239,400 on recombinant antibodies and their production; WO 1991/09967 on humanized antibodies; WO 1992/01047 on antibody production; WO 1992/22653 on methods for making humanized antibodies; WO 1993/11161 on multivalent antigen-binding proteins; WO 1994/13804 on multivalent antigen-binding proteins; WO 2000/66631 on specific binding members for TGF-beta; and Henry “Special Delivery: Alternative methods for delivering drugs improve performance, convenience, and patient compliance.” C&EN, p. 49-65 (2000). See also U.S. Pat. Nos. 6,140,471 and 5,969,108 and 5,872,215 and 5,871,907 and 5,858,657 and 5,837,242 and 5,733,743; EP 1,024,191; EP 774,511; WO 1997/13844; EP 656,941and 605,522 and WO 1994/13804; EP 589,877; EP 585,287; WO 1993/19172; EP 540,586; WO 1993/06213; WO 1992/20791; WO 1992/01787; and WO 1992/01047. Further, WO 2004/065417 discloses various alterations to antibodies and antigen-binding fragments to improve yield. See also US 20050049403.
There is a need to control TGF-beta molecules to prevent their deleterious effects in diseases such as those set forth above. There is also a need to provide monoclonal antibodies of high affinity that bind to TGF-beta specifically and that neutralize TGF-beta activity so as to act as a TGF-beta antagonist. The apparent loss of TGF-beta regulation by neoplastic cells coupled with the suppression of immune function and the TGF-beta-induced stroma formation makes potential intervention with TGF-beta antagonists an attractive option for cancer therapy. In addition, TGF-beta antibodies are useful in diagnostic assays and immunoaffinity purification.